The clinical presentations of many important ocular infections have significant overlap, and thus it is often difficult to establish the correct etiological diagnosis on purely clinical grounds. Recently, we have developed an enzyme immunofiltration staining method for rapid laboratory diagnosis of ocular viral and chlamydial infections. We propose to perfect this method so that it may be used in a clinical setting to detect herpes simplex virus type 1 (HSV-1) herpes simplex virus type 2 (HSV-2) varicella zoster virus (VZV), and chlamydia trachomatic antigens directly in specimens obtained from ocular lesions. Preliminary studies with HSV, VZV and chlamydia have shown that this method is rapid enough (10 minutes) so the results are available in time to affect therapeutic decisions. We propose to continue our optimization of this method in HSV, VZV and chlamydial infections by selecting pools of monoclonal antibodies which are capable of detecting all virus strains or chlaymdial serovars and testing for methods of inactivating native peroxdidases. We will develop the differential bead immunoassay system of detecting soluble viral antigens and perfect this technique and the reagents needed to extend its use to the rapid diagnosis of ocular infections by adenovirus (AV) and enterovirus 70 (EV-70). We also propose to use the immunofiltration technique to detect and quantitate IgA, IgG, IgE and IgM antibodies to HSV and VZV in serial samples of tear fluid and serum from infected patients. The results of these studies will be correlated with data on agent isolation, antigen detection, clinical manifestations, response to therapy, and outcome in order to determine which, if any, of these local or humoral, agent specific, responses have prognostic significance. We also propose to do identical studies in the rabbit model system where we have already followed 20 rabbits for over 4 years and have collected thousands of specimens.